雷帕霉素对人淋巴瘤Raji细胞增殖、VEGF-A及HIF-1α表达的影响

摘要

Objective:To investigate the influence of rapamycin to proliferation of human lymphoma cell line Raji cells in vitro,the expression of VEGF-A and HIF-1α,and discuss its clinical significance.Methods:After Raji cells treated with rapamycin at different concentrations (0,10,50,100,250,500nmol/L)and different time (24, 48,72h),its effects on cell proliferation were observed by CCK8 method.Its effects on mTOR,VEGF,protein HIF-1αwere detected by using Western blot(WB) in 72h.Its effects on mRNA were detected by using Reverse transcription (RT-PCR).Results:Rapamycin had inhibitory effect on Raji cells proliferation obviously (P ＜ 0.01).Raji cells were treated with rapamycin after 72h,with the increase of drug concentration,the protein of mTOR,VEGF and HIF-1 α were decreased significantly (P ＜ 0.01).After Raji cells were treated with rapamycin,with the increase of drug concentration and the increase of treatment time,the mRNA of mTOR,VEGF and HIF-1 α were decreased obviously (P ＜ 0.01).The expression of mRNA in mTOR,VEGF and protein HIF-1α had a positive correlation between each pairs.Conclusion:Rapamycin have an inhibitory effect in human lymphoma Raji ceils though mTOR pathway obviously,can reduce the translation of mRNA of mTOR,VEGF-A,HIF-1α apparently,and reduce their transcription of protein to inhibit the growth of Raji cells and their angiogenesis.%目的:探讨雷帕霉素对人淋巴瘤细胞株Raji细胞体外增殖、VEGF-A及HIF-1α表达的影响及其临床意义.方法:不同浓度(0、10、50、100、250、500nmol/L)雷帕霉素对Raji细胞作用不同时间(24、48、72h)后,使用CCK8法检测细胞增殖的变化;采用蛋白质印记法(Western blot,WB)检测72h组不同浓度mTOR、VEGF-A、HIF-1α蛋白变化;采用反转录聚合酶链式反应(RT-PCR)检测其mRNA的变化.结果:雷帕霉素对Raji细胞增殖具有显著抑制作用(P＜0.01);雷帕霉素处理72h后的Raji细胞,随着药物浓度增加,mTOR、VEGF-A和HIF-1 α蛋白明显减少(P＜0.01);雷帕霉素处理后的Raji细胞,随着药物浓度和作用时间的增加,mTOR、VEGF-A和HIF-1α的mRNA明显减少(P＜0.01);mTOR、VEGF-A及HIF-1α的mR-NA表达两两之间呈正相关性.结论:雷帕霉素通过mTOR通路,能明显抑制人淋巴瘤Raji细胞增殖过程,表现出明显的时间、剂量效应依赖关系;同时也可明显减少mTOR、VEGF-A、HIF-1α的mRNA的转录表达,减少p-mTOR、VEGF-A、HIF-1α等蛋白的翻译表达,从而抑制Raji细胞的生长及其血管新生.